Workflows for Glycosylation and Sialic Acid Analysis of Biotherapeutic Glycoproteins

N-Glycans on biotherapeutics can affect immunogenicity, pharmacokinetics and pharmacodynamics, making characterization of N-glycans essential in the development process. Here we present rapid N-glycan sample preparation and analysis workflows for biotherapeutics, including released N-glycan labeling with InstantPC and 2-AB for characterization by liquid chromatography (LC) with fluorescence (FLD) and mass spec (MS) detection. In addition, we will discuss approaches to identify different sialylated N-glycan species as well as quantitating the sialic acid content of biotherapeutic glycoproteins using plate-based and LC/FLD/MS approaches.